Method for separation of anionic oligosaccharides

• US 5,145,956 A
• Filed: 12/11/1991
• Issued: 09/08/1992
• Est. Priority Date: 12/11/1991
• Status: Expired due to Fees

• Current Assignee: Glycomed, Inc. (DSM-Firmenich AG)
• Original Assignee: Glycomed, Inc. (DSM-Firmenich AG)
• Inventors: Tyrrell, David, Lam, Lun H.
• Primary Examiner(s): GRIFFIN, RONALD
• Application Number:
  US07/805,501
• Time in Patent Office: 272 Days
• Field of Search: 536/124, 536/127, 436/807, 436/809
• US Class Current: 536/124
• CPC Class Codes: B01J 20/3272 Polymers obtained by reactions otherwise than involving only carbon to carbon unsaturated bonds
• CPC Class Codes: C07H 1/08 from natural products
• CPC Class Codes: C08B 37/0078 Degradation products
• CPC Class Codes: Y10S 436/807 Apparatus included in process claim, e.g. physical support structures
• CPC Class Codes: Y10S 436/809 Multifield plates or multicontainer arrays

Abstract

Separation of anionic oligosaccharides on a preparative scale with high resolution can be achieved by anionic exchange chromatography using PEI-derivatized supports.

Claims

1. A method to separate components of a mixture of anionic oligosaccharides lacking carbonyl groups, which method comprises:

applying said mixture to a chromatographic column wherein said chromatographic column contains a solid support coated with polyethyleneimine (PEI) under conditions where said anionic oligosaccharides are adsorbed to the column;

followed by eluting said column with a salt gradient to obtain a multiplicity of fractions so as to separate said anionic oligosaccharide components.

2. The method of claim 1 wherein said mixture contains anionic oligosaccharides of substantially the same molecular weight.

3. The method of claim 1 wherein said anionic oligosaccharides are obtained by depolymerization of a glycosaminoglycan.

4. The method of claim 3 wherein said glycosaminoglycan is heparin.

5. The method of claim 1 wherein said salt gradient elution is conducted using a lyophilizable salt.

6. The method of claim 5 wherein said salt is ammonium bicarbonate.

7. The method of claim 1 wherein said anionic oligosaccharides have been prepared by reduction of carbonyl-containing oligosaccharides with a metal hydride.

8. A method to modify a chromatographic support coated with polyethyleneimine (PEI), which method comprises treating said support with a low molecular weight aldehyde in an amount and for a time sufficient to block primary amino groups contained in said PEI, followed by removing excess aldehyde from the support.

9. A chromatographic support prepared by the method of claim 8.

10. A method to separate components of a mixture of anionic oligosaccharides at least some of which contain carbonyl groups, which method comprises:

applying said mixture to a chromatographic column wherein said chromatographic column contains a solid support coated with polyethyleneimine (PEI) modified by the method of claim 8 under conditions where said anionic oligosaccharides are adsorbed to the column;

followed by eluting said column with a salt gradient to obtain a multiplicity of fractions so as to separate said anionic oligosaccharide components.